Title:
NITRILASE MUTANT AND APPLICATION THEREOF IN PREPARATION OF 1-CYANOCYCLOHEXANEACETIC ACID
Document Type and Number:
WIPO Patent Application WO/2021/169490
Kind Code:
A1
Abstract:
Provided is a nitrilase mutant and application thereof in the preparation of 1-cyanocyclohexaneacetic acid, the mutant being obtained by mutating one or more of the 180th and 205th amino acids in the amino acid sequence shown in SEQ ID No.2. By means of semi-rational design and protein molecular modification, the specific enzyme activity of a nitrilase double mutant AcN-G180D/A205C is up to 1.6 times higher, and the conversion rate is higher than 99%. In addition, the use of recombinant E. coli containing the nitrilase mutant to hydrolyze 1-cyanocyclohexylacetonitrile at a high temperature (50°C) shortens the reaction time to a quarter of the original, such that the obtained mutant has a broad application prospect in the efficient catalysis of 1-cyanocyclohexylacetonitrile to synthesize an intermediate, 1-cyanocyclohexaneacetic acid, for Gabapentin.
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Inventors:
XUE YAPING (CN)
XIONG NENG (CN)
LI QIAN (CN)
ZHENG YUGUO (CN)
XIONG NENG (CN)
LI QIAN (CN)
ZHENG YUGUO (CN)
Application Number:
PCT/CN2020/135582
Publication Date:
September 02, 2021
Filing Date:
December 11, 2020
Export Citation:
Assignee:
UNIV ZHEJIANG TECHNOLOGY (CN)
International Classes:
C12N1/21; C12N9/78; C12N15/55; C12N15/70; C12P13/00
Domestic Patent References:
| WO2006069110A2 | 2006-06-29 |
Foreign References:
| CN111471668A | 2020-07-31 | |||
| CN108486088A | 2018-09-04 | |||
| CN107177576A | 2017-09-19 | |||
| CN102911975A | 2013-02-06 | |||
| CN101629192A | 2010-01-20 | |||
| CN104212784A | 2014-12-17 |
Other References:
LIU, Z.Q. ET AL.: "Screening and Improving the Recombinant Nitrilases and Application in Biotransformation of Iminodiacetonitrile to Iminodiacetic Acid", PLOS ONE, vol. 8, no. 6, 27 June 2013 (2013-06-27), XP055632049, DOI: 10.1371/journal.pone.0067197
Attorney, Agent or Firm:
TIANZHENG PATENT ATTORNEYS (CN)
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